The South African Breweries Section : 12.

32
Page : 1 of 2

BREWING MANUAL Revision : Draft Rev

Date : Aug 2002
Vol. VIII – Microbiological Methods
Section 12: Techniques:
12.32 – Inoculation: Spread Plate technique

1 INTRODUCTION b) Remove a sterile pipette from its wrapping

or canister and draw the tip through the
This method of inoculation onto solid media flame.
is designed to detect a low level of c) Mix the contents of the sample bottle or
contamination in a sample. tube.
For example, culture yeast. d) Remove the lid or cap, flame the mouth and
The relatively high volume of inoculum withdraw between 0,1 ml and 1,0 ml of
allows for increased sensitivity. sample into the pipette, as required.
e) Replace the lid or cap.
2 HEALTH AND SAFETY f) Lift the lid of the petri dish slightly, and
expel the sample onto the surface of the
Section 3 Agar.
g) Sterilise the Drigalski spatula by flash-
3 REFERENCES flaming with 96% alcohol.
h) Cool the Drigalski spatula on the edge of the
Media: section 9 Agar.
i) Spread the sample over the surface, rotating
4 MEDIA AND REAGENTS the plate to ensure even distribution.
j) Replace the lid of the petri dish.
Plates of solid media (section 9)
Notes:
Physiological saline (MMM: 9.11)
i. Drigalski
spatulas will not have an indefinite
5 APPARATUS
lifespan.
a) Laminar flow bench
ii. Examine weekly
b) Bunsen burner
for tiny cracks, especially when frequent
c) Cotton wool swabs
inoculation of spread plates is carried
d) Sterile pipettes
out.
e) Sample to be inoculated
iii. Spores may
f) Drigalski spatula
attach to the fissures and
cross-contaminate plates.
6 PROCEDURE

a) Aseptic technique must be observed